Fluorescence Lifetime Imaging

XiaoTao Hao, Lachlan McKimmie, Allison Heskes

In addition to being able to measure the fluorescence lifetimes of bulk solutions or films, using a scanning microscope we are also able to generate lifetime images of fluorescent samples. These images give additional information over intensity based images as a fluorescence decay profile is collected for every pixel. Examining the changes in fluorescence lifetimes across the image aids in determining the local environments of the fluorophores. The image below is of a wool fibre; here the different colours represent different fluorescent lifetimes.

FLIM-Wool-Fibre1

Relevant papers:

  • T.A. Smith, C.N. Lincoln and D.K. Bird, “Time-Resolved Fluorescence in Microscopy” in Fluorescence Applications in Biotechnology and Life Sciences, E.M. Goldys (Ed.), Wiley-Blackwell, ISBN 978-0-470-08370-3, pp 195-221 (2009)
  • D.K. Bird, A.L. Schneider, A. Watkinson, B. Finnin and T.A. Smith, “Navigating Transdermal Diffusion with Multiphoton Fluorescence Lifetime Imaging”, J. Microscopy, (2008), 230(1), 61-69
  • D.K. Bird, K.M. Agg, N.W. Barnett and T.A. Smith, “Time-Resolved Fluorescence Microscopy of Gunshot Residue: An Application to Forensic Science”, J. Microscopy, 226(Pt 1), 18-25 (2007)
  • S. Mathai, D.K. Bird, S.S. Stylli, T.A. Smith and K.P. Ghiggino, “Two-photon Absorption Cross-Sections and Time-Resolved Fluorescence Imaging Using Porphyrin Photosensitisers”, Photochem. Photobiol. Sci., (2007), 6, 1019-1026